2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38561Hearing loss, caused by gene mutations and environmental factors, is a common sensory disorder in the human population. In the developed countries, the incidence of congenital hearing loss is estimated at 1 in 1000 births, of which approximately 60% cases are attributed to genetic factors. To date, 59 auditory genes have been identified. Previously, we have found many mutations in the Cx29 and CLDN14 from screening of 260 patients with nonsyndromic deafness. However, functional alteration in these mutant genes remain unknown. The overall goal of this project is to study the pathogenesis of Cx29 and CLDN14 from non-syndromic deafness in Taiwan. Simultaneously, we will also be interested in uncovering the functional interaction between tight junction gene CLDN14 and gap junction gene, Cx26, Cx30 and Cx29. In the proposed three-year project, we will focused on study the functional consequence of these genes when mutations occur and functional interaction of tight junction gene CLDN14 with gap junction gene of Cx26, Cx30 and Cx29, using approaches of molecular biological, cell biological and electron microscopy. Firstly, we will investigate their functional properties and interaction amongst these mutant proteins expressed in HeLa or MDCK cell including establishing stable cell line, expressing products of both wild type and mutant forms of these genes. Based on previous study and results from the first year, we will study functional interaction of these genes with Cx26 or Cx30 in the second year. Finally, we will to study functional interaction between Cx29 and CLDN14 gene in HeLa or MDCK cell. Further more, we will use cochlear cell line (Stria Vascularis SV-K1) to study functional interaction of tight junction gene CLDN14 and gap junction gene, Cx26, Cx30 and Cx29. We anticipate that the study will further our understanding of the functional role of these genes in nonsyndromic deafness. We believe that the etiology and mechanism of pathogenesis in hereditary hearing loss will be comprehensively appreciated upon completion of this research.聽障可因遺傳基因突變或環境因素，或兩者兼之引起的。約有千分之一嬰兒在出生時罹患重度聽障，在已開發國家約有60%個案是遺傳因素，目前已知有59個基因的突變會導致聽障，各基因的致病機制不盡相同，非常複雜。在我們先前對260位非症候群聽障患者的篩檢研究中，我們發現在非症候群聽障患者CONNEXIN29(Cx29)和CLAUDIN14 (CLDN14)基因帶有突變點。然而目前對於這些基因發生突變後所產生的突變蛋白對功能的影響和是否會影響到其他基因蛋白質的功能尚未清楚，且同一基因在不同位置的突變常會產生功能上的差異。因此本計畫的研究動機是探討Cx29和CLDN14基因突變後對其功能的影響和彼此之間所造成的影響。另外也將探討這兩基因和Cx26及Cx30基因之間的交互作用，以瞭解與K+ 再循環(recycling)和維持耳蝸內K+濃度相關基因在聽障中的致病機制。 本計畫中我們將針對Cx29和CLDN14基因突變後在非症候群聽障的功能影響或此兩基因和Cx26及Cx30基因之間的交互作用或此兩基因彼此之間的交互作用，利用分子生物學、細胞生物學、電子顯微鏡等研究法來加以探討。本計劃分三年來執行，第一年將分別以HeLa細胞和MDCK細胞來探討單基因(Cx29和CLDN14)突變後造成功能上的影響並建立穩定細胞表現株，第二年由於Cx26和Cx30是耳蝸內表現豐富的蛋白，因此我們將以建立好的穩定細胞表現株和老鼠耳蝸組織來探討Cx29或CLDN14基因分別與Cx26和Cx30基因的交互作用。第三年以HeLa細胞和MDCK細胞來探討Cx29和CLDN14之間的交互作用，並以耳蝸細胞的細胞株(Stria Vascularis SV-K1)來研究gap junction系統(Cx26、Cx30和Cx29)和tight junction系統(CLDN14)之間的相互關係。相信這三年計畫如能獲得支持將對於多種聽障基因之作用機轉的瞭解將有助於幫助我們釐清造成非症候群聽障的真正致病機轉。Cx29CLDN14非症候群聽障功能研究Cx29CLDN14non-syndromic deafnessfunctional studyProject